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Strengthening regional stem cells based research for advancement of multi modal innovative strategy for modelling neurodevelopmental disorders (STREAMLINE)
Get PDFNeurodevelopmental disorders (NDDs) caused by alterations in early brain development are a
group of complex and heterogeneous disorders that give rise to the psychiatric conditions such
as autism spectrum disorders, intellectual disability, schizophrenia and bipolar disorder. NDDs
have a high socio-economic impact and patients with NDDs and mental health problems are
geographically dispersed. Thus, establishment of high capacity regional centres with equivalent
research capacity and codes of practice is needed in order to build global research activity. The
goals of STREAMLINE are to enhanced strategic networking and reinforce research and
innovation potential of the Institute of Molecular Genetics and Genetic Engineering (IMGGE) in
order to develop IMGGE as a high capacity hub for research of NDDs in the Western Balkans
which is able to integrate with similar centres in Europe. This will be achieved by twinning
IMGGE with three top-class research institutions in Europe: Cardiff University, University of
Maastricht and Centre for Research and Technology. The project objectives are: a) to enhance
the current and scale up the overall strategic networking activities between the IMGGE and
internationally-leading European partners and regional partners; b) to raise the research profile
of research stuff and scientific attractiveness of IMGGE in the field of NDDs; c) to strengthens
research management capacities and administrative skills of the IMGGE staff and d) to become
a regional leader in innovations tackling NDDs. Objectives will be achieved through networking,
staff exchanges, training, summer schools, symposia, workshops, participation at conferences
and dissemination in scientific community, industries and public. Twinning with partners will
provide access to new research avenues, creativity, development of new approaches and
increased mobility (inwards and outwards) of qualified scientists.Principal Investigator: Academician Milena StevanovicDuration period: 2022-202
Mechanisms of desiccation tolerance in Ramonda serbica Panc.: transcriptomic, proteomic, metabolomic, and photosynthetic aspects
Get PDFRamonda serbica Panc. is a resurrection plant species that can survive desiccation for a long
period and fully resume metabolic functions upon rewatering in a very short period, even within
48 h. The goal of this study was to identify key candidates and pathways involved in desiccation
tolerance in R. serbica. To achieve this, systems biology approach combining transcriptomics,
proteomics, and analysis of specific metabolites was employed. In addition, FTIR analysis of the
cell wall polymers and a detailed analysis of the photosynthetic electron transport (PET) chain
were performed. In total, 1192 different protein groups were quantified by TMT-based comparative
quantitative proteomics. Among them, 408 protein groups showed a statistically significant
difference in abundance between hydrated (HL) and desiccated leaves (DL). Upon desiccation, the
majority of proteins related to photosynthetic processes were less abundant, while chlorophyll
fluorescence measurements implied shifting from linear photosynthetic transport (PET) to cyclic
electron transport (CET). The amounts of H2O2 scavenging enzymes, including ascorbate-glutathione
cycle components, catalases, peroxiredoxins, Fe-, and Mn- superoxide dismutase (SOD) were
reduced in DL. However, four Cu/ZnSOD isoforms, three polyphenol oxidases, six germin-like proteins
(GLPs), and 22 late embryogenesis abundant proteins (LEAPs; mainly LEA4 and dehydrins),
were desiccation-inducible. Desiccation-induced cell wall remodelling by changes in cell wall
polymer composition might be linked with pectin demethylesterification and GLP-derived H2O2/
HO•. Our study demonstrated that desiccation tolerance in R. serbica is a complex, species-specific
process orchestrated by several metabolic pathways and regulatory networks acting at the transcript,
protein, metabolite and physiological levels.Book of Abstracts: 4th International Conference on Plant Biology and 23rd SPPS Meeting, 6-8 October 2022, Belgrad
Role of hsa-miR-93-5p in colorectal cancer
Get PDFKolorektalni karcinom predstavlja heterogenu bolest koja zauzima treće i drugo mesto na svetu po stepenu
incidence i stopi mortaliteta, respektivno. Poznato je da mikroRNK molekuli imaju ključne uloge u
procesu patogeneze, progresije i metastaziranja kolorektalnog karcinoma, ali i rezistenciji na hemioterapiju.
U različitim tumorima je pokazano da hsa-miR-93-5p može imati onkogenu, ali i tumor-supresorsku
ulogu. Ovaj rad ima za cilj da predstavi dosadašnja saznanja o ulozi hsa-miR-93-5p u procesima vezanim
za kolorektalnu karcinogenezu, metastaziranje i odgovor na hemioterapiju kod pacijenata sa kolorektalnim
karcinomom. Rad takođe razmatra hsa-miR-93-5p kao potencijalni prognostički i prediktivni biomarker
i primenu ovog molekula u kliničkoj praksi.Colorectal cancer is a heterogeneous disease that ranks third and second globally in terms of incidence
and mortality rates, respectively. MicroRNAs have been implicated in the growth, progression, metastasis
and response to therapy of different tumors, including colorectal cancer. Hsa-miR-93-5p has been
reported to have oncogenic and tumor-suppressive roles in different tumor kinds. This review article aims
to present the current knowledge on the role of hsa-miR-93-5p in the processes related to colorectal carcinogenesis,
metastasis and response to chemotherapy in patients with primary and metastatic colorectal
cancer. Also, the role of hsa-miR-93-5p as a potential prognostic and predictive biomarker and
application of this molecule in the clinical practice is described.Posebno se zahvaljujem dr Aleksandri Nikolić na kritičkoj oceni ovog rada, kao i svim savetima i sugestijama za poboljšanje njegovog kvaliteta
Chemical profile and antioxidative activity of red wines obtained from autochthonous and international grape clone varieties
Get PDFOva tema je fokusirana na hemijsku i biološku karakterizaciju odabranih uzoraka crvenih vina. Akcenat
je stavljen na hemijsku karakterizaciju klonskih sortnih vina vranac i merlo, kao i na procenu antioksidativne
aktivnosti merlo vina u ćelijama kvasca. U radu su primenjene brojne analitičke tehnike za
određivanje koncentracije fenolnih jedinjenja, elementalnog sastava i in vitro antioksidativnu aktivnosti
ispitivanih uzoraka (spektrofotometrija, ciklična voltametrija, klasična polarografija, EPR spektroskopija,
LC-MS/MS, ICP-OES i ICP-MS). Pored antioksidativne, ispitane su i citotoksična i antibakterijska aktivnost odgovarajućih
uzoraka vina. Eksperiment na bazi vinskog kvasca je obuhvatio određivanje ukupnih –SH
grupa, kao i enzimskih aktivnosti glutation peroksidaze, glutation reduktaze i katalaze. Eksperimentalno
dobijeni podaci za klonska sortna vina merlo ukazuju na poseban doprinos glutation peroksidaze preživljavanju
ćelija kvasca u uslovima oksidativnog stresa. Konačno, primenom veštačke neuronske mreže razvijena
je brza i precizna metoda za predviđanje uticaja malih promena u vinskom fenolnom profilu na
aktivnosti istaknutih enzima antioksidativne zaštite.This topic focuses on the chemical and biological characterization of selected red wine samples. Emphasis
is placed on the chemical characterisation of clonal wines of Vranac and Merlot varieties, as well as
on the evaluation of the antioxidant activity of Merlot wines in yeast cells. Numerous analytical techniques
were used to determine the concentration of phenolic compounds, elemental composition and in
vitro antioxidant activity of the tested samples (spectrophotometry, cyclic voltammetry, classical polarography,
EPR spectroscopy, LC-MS/MS, ICP-OES and ICP-MS). In addition, total antioxidant activity, cytotoxic
and antibacterial activity of the respective wine samples were determined. The wine yeast experiment
involved determining the total groups –SH, as well as enzymatic activities of glutathione peroxidase, glutathione
reductase and catalase. The experimentally obtained data for clonal Merlot wine varieties indicate
a particular contribution of glutathione peroxidase to the survival of yeast cells under oxidative stress conditions.
Finally, using an artificial neural network (ANN), a rapid and accurate method was developed to predict
the effect of small changes in the wine phenolic profile on the activities of prominent antioxidant
enzymes
Ретке болести у ери геномике
Get PDFTokom poslednjih decenija, istraživanja na polju retkih bolesti napreduju
ogromnom brzinom usled sve veće upotrebe sekvenciranja nove generacije (engl.
Next-generation sequencing, NGS). Sekvenciranje kompletnog ljuskog genoma (engl.
Whole genome sequencing, WGS) osoba koje boluju od retkih bolesti je postalo
lako dostupno. Pored pronalaženja novih varijanti i novih gena koji leže u
osnovi retkih bolesti, genomika je omogućila i otkriće gena modifikatora koji
mogu da objasne uočene nedoslednosti u korelaciji genotipa i fenotipa.
Fenilketonurija je urođena metabolička retka bolest koja je uzrokovana
varijantama u genu za fenilalanin hidroksilazu (PAH). U ovoj studiji, sproveli
smo sekvenciranje kompletnog genoma 4 osobe iz nepovezanih nesrodnih porodica
koje su imale patogene varijante u PAH genu, ali nisu razvile fenilketonuriju
uprkos tome što nisu bile lečene. Otkrili smo dve nove varijante, p.Pro1591Ala
u SHANK1 i p.Asp18Asn u SHANK2 genima, kao i prethodno opisane
SHANK2:p.Gly46Ser, SHANK2:p.Pro1388_Phe1389insLeuPro i SHANK3:p.Pro1716Thr
varijante. Računarske predikcije su pokazale da identifikovane varijante ne
ukidaju funkciju SHANK proteina. Međutim, promene u posttranslacionim
modifikacijama SHANK proteina mogu uticati na funkcionisanje
glutamatergičnih sinapsi, regulaciju citoskeleta i doprineti održavanju
optimalne sinaptičke gustine i broja dendritskih bodlji. Naši rezultati po
prvi put povezuju porodicu SHANK gena i osobine neuroloških promena kod
osoba sa fenilketonurijom.Током последњих деценија, истраживања на пољу ретких болести напредују
огромном брзином услед све веће употребе секвенцирања нове генерације (енгл.
Next-generation sequencing, NGS). Секвенцирање комплетног љуског генома (енгл.
Whole genome sequencing, WGS) особа које болују од ретких болести је постало
лако доступно. Поред проналажења нових варијанти и нових гена који леже у
основи ретких болести, геномика је омогућила и откриће гена модификатора који
могу да објасне уочене недоследности у корелацији генотипа и фенотипа.
Фенилкетонурија је урођена метаболичка ретка болест која је узрокована
варијантама у гену за фенилаланин хидроксилазу (PAH). У овој студији, спровели
смо секвенцирање комплетног генома 4 особе из неповезаних несродних породица
које су имале патогене варијанте у PAH гену, али нису развиле фенилкетонурију
упркос томе што нису биле лечене. Открили смо две нове варијанте, p.Pro1591Ala
у SHANK1 и p.Asp18Asn у SHANK2 генима, као и претходно описане
SHANK2:p.Gly46Ser, SHANK2:p.Pro1388_Phe1389insLeuPro и SHANK3:p.Pro1716Thr
варијанте. Рачунарске предикције су показале да идентификоване варијанте не
укидају функцију SHANK протеина. Међутим, промене у посттранслационим
модификацијама SHANK протеина могу утицати на функционисање
глутаматергичних синапси, регулацију цитоскелета и допринети одржавању
оптималне синаптичке густине и броја дендритских бодљи. Наши резултати по
први пут повезују породицу SHANK гена и особине неуролошких промена код
особа са фенилкетонуријом.Knjiga sažetaka: Treći Kongres biologa Srbije, Zlatibor, Srbija 21 - 25. 9. 2022
Supplementary data for the article: Stevanović, M. M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J., & Boccaccini, A. R. (2022). Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. Journal of Biomaterials Applications, 36(10), 1800–1811. [https://doi.org/10.1177/08853282211073731]
Get PDFThis is supplementary material for paper: Stevanović, M. M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J., & Boccaccini, A. R. (2022). Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. Journal of Biomaterials Applications, 36(10), 1800–1811. https://doi.org/10.1177/08853282211073731Related to published version:[https://imagine.imgge.bg.ac.rs/handle/123456789/1773
Comparative genomics of trimethoprim-sulfamethoxazole-resistant Achromobacter xylosoxidans clinical isolates from Serbia reveals shortened variant of class 1 integron integrase gene
No full textTrimethoprim-sulfamethoxazole (SXT) is the preferable treatment option of the infections caused by Achromobacter spp. Our study aimed to analyze the SXT resistance of 98 Achromobacter spp. isolates from pediatric patients, among which 33 isolates were SXT-resistant. The presence of intI1 was screened by PCR and genome sequence analyses. The intI1 gene was detected in 10 of SXT-resistant isolates that had shorter intI1 PCR fragments named intI1S. Structural changes in intI1S were confirmed by genome sequencing and analyses which revealed 86 amino acids deletion in IntI1S protein compared to canonical IntI1 protein. All IntI1S isolates were of non-CF origin. Pan-genome analysis of intI1S bearing A. xylosoxidans isolates comprised 9052 genes, with the core genome consisting of 5455 protein-coding genes. Results in this study indicate that IntI1S isolates were derived from clinical settings and that cystic fibrosis (CF) patients were potential reservoirs for healthcare-associated infections that occurred in non-CF patients
Biocompatibility and antibiofilm activity of graphene-oxide functionalized titanium discs and collagen membranes
No full textThe aims of the study were: 1) to evaluate the effect on biofilm formation of barrier membranes and titanium surfaces coated with graphene-oxide (GO); 2) to analyze the connection between the superficial topography of the tested materials and the amount of bacterial accumulation on them and 3) to analyze the biocompatibility of GO functionalized discs using the zebrafish model. Methods: Single species bacterial biofilms (Streptococcus oralis, Veilonella parvula, Fusobacterium nucleatum, Porphyomonas gingivalis) were grown on GO-free membranes, membranes coated with 2 and 10 mu g/ml of GO, GO-free and GO-coated titanium discs. The biofilms were analyzed by determining the CFU count and by Scanning Electron Microscopy (SEM) and the materials' topography by Atomic Force Microscopy (AFM). Zebrafish model was used to determine the materials' toxicity and inflammatory effects. Results: AFM showed similar roughness of control and GO-coated materials. CFU counts on GO-coated discs were significantly lower than on control discs for all species. CFU counts of S. oralis, V. parvula and P. gingivalis were lower on biofilms grown on both types of GOcoated membranes than on GO-free membrane. SEM analysis showed different formation of single species biofilm of S. oralis on control and GO-coated materials. GO-functionalized titanium discs do not induce toxic or inflammatory effects. Significance: Titanium implant surfaces functionalized with GO have shown to be biocompatible and less susceptible to biofilm formation. These results encourage further in vivo investigation of the tested materials on infection prevention, specifically in prevention and reduction of peri-implant mucositis and periimplantitis incidence
Streptomyces sp. BV410: Interspecies cross-talk for staurosporine production
No full textAims Sequencing and genome analysis of two co-isolated streptomycetes, named BV410-1 and BV410-10, and the effect of their co-cultivation on the staurosporine production. Methods and Results Identification of two strains through genome sequencing and their separation using different growth media was conducted. Sequence analysis revealed that the genome of BV410-1 was 9.5 Mb, whilst that of BV410-10 was 7.1 Mb. AntiSMASH analysis identified 28 biosynthetic gene clusters (BGCs) from BV410-1, including that responsible for staurosporine biosynthesis, whilst 20 BGCs were identified from BV410-10. The addition of cell-free supernatant from BV410-10 monoculture to BV410-1 fermentations improved the staurosporine yield from 8.35 mg L-1 up to 15.85 mg L-1, whilst BV410-10 monoculture ethyl acetate extract did not have the same effect. Also, there was no improvement in staurosporine production when artificial mixed cultures were created using three different BV410-1 and BV410-10 spore ratios. Conclusions The growth of BV410-10 was inhibited when the two strains were grown together on agar plates. Culture supernatants of BV410-10 showed potential to stimulate staurosporine production in BV410-1, but overall co-cultivation attempts did not restore the previously reported yield of staurosporine produced by the original mixed isolate. Significance and Impact of Study This work confirmed complex relations between streptomycetes in soil that are difficult to recreate under the laboratory conditions. Also, mining of streptomycetes genomes that mainly produce known bioactive compounds could still be the fruitful approach in search for novel bioactive molecules
Strong Antibiotic Activity of the Myxocoumarin Scaffold in vitro and in vivo
Get PDFThe increasing emergence of resistances against established antibiotics is a substantial threat to human health. The discovery of new compounds with potent antibiotic activity is thus of utmost importance. Within this work, we identify strong antibiotic activity of the natural product myxocoumarin B from Stigmatella aurantiaca MYX-030 against a range of clinically relevant bacterial pathogens, including clinical isolates of MRSA. A focused library of structural analogs was synthesized to explore initial structure-activity relationships and to identify equipotent myxocoumarin derivatives devoid of the natural nitro substituent to significantly streamline synthetic access. The cytotoxicity of the myxocoumarins as well as their potential to cure bacterial infections in vivo was established using a zebrafish model system. Our results reveal the exceptional antibiotic activity of the myxocoumarin scaffold and hence its potential for the development of novel antibiotics